Plants have evolved an amazing array of metabolic pathways leading to molecules capable of responding promptly and effectively to stress situations imposed by biotic and abiotic factors, some of which supply the ever-growing needs of humankind for natural chemicals, such as pharmaceuticals, nutraceuticals, agrochemicals, food and chemical additives, biofuels, and biomass. In Plant Secondary Metabolism Engineering: Methods and Applications, expert researchers provide detailed practical information on some of the most important methods employed in the engineering of plant secondary metabolism pathways and in the acquisition of essential knowledge in performing this activity, including the significant advances and emerging strategies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Plant Secondary Metabolism Engineering: Methods and Applications will aid scientists engaged in the challenging task of modifying some of the most intricate products of plant evolution and support their efforts directed toward the vital goal of sustainable natural chemicals.

1. Plant Secondary Metabolism and Challenges in Modifying Its Operation: An Overview Naíla Cannes do Nascimento and Arthur Germano Fett-Neto 2. Suppression Subtractive Hybridization as a Tool to Identify Anthocyanin Metabolism-Related Genes in Apple Skin Yusuke Ban and Takaya Moriguchi 3. Identification of Regulatory Protein Genes Involved in Alkaloid Biosynthesis Using a Transient RNAi System Yasuyuki Yamada, Nobuhiko Kato, Yasuhisa Kokabu, Qingyun Luo, Joseph Gogo Dubouzet, and Fumihiko Sato 4. Site-Directed Mutagenesis and Saturation Mutagenesis for the Functional Study of Transcription Factors Involved in Plant Secondary Metabolite Biosynthesis Sitakanta Pattanaik, Joshua R. Werkman, Que Kong, and Ling Yuan 5. Isolation of Proteins Binding to Promoter Elements of Alkaloid Metabolism-Related Genes Using Yeast One-Hybrid Debora Vom Endt and Giancarlo Pasquali 6. Modulation of Carotenoid Accumulation in Transgenic Potato by Inducing Chromoplast Formation with Enhanced Sink Strength Joyce Van Eck, Xiangjun Zhou, Shan Lu, and Li Li 7. Overexpression of Rate-Limiting Enzymes to Improve Alkaloid Productivity Tomoya Takemura, Yit-lai Chow, Takehiko Todokoro, Takuya Okamoto, and Fumihiko Sato 8. Microbial Expression of Alkaloid Biosynthetic Enzymes for Characterization of Their Properties Hiromichi Minami, Nobuhiro Ikezawa, and Fumihiko Sato 9. Producing a Recombinant Flavin-Containing Monooxygenase from Coffea arabica in Escherichia coli for Screening of Potential Natural Substrates Igor Cesarino and Paulo Mazzafera 10. Efficient Production of Active Form Recombinant Cassava Hydroxynitrile Lyase Using Escherichia coli in Low-Temperature-Culture Hisashi Semba, Eita Ichige, Tadayuki Imanaka, Haruyuki Atomi, and Hideki Aoyagi 11. Introduction of the Early Pathway to Taxol Biosynthesis in Yeast by Means of Biosynthetic Gene Cluster Construction Using SOE-PCR and Homologous Recombination Pia Dahm and Stefan Jennewein 12. Biocatalytic Synthesis of Tritium (3H) Labelled Taxa-4(5),11(12)-diene, the Pathway Committing Precursor of the Taxoid Diterpenoids Hans Schmeer and Stefan Jennewein 13. USER Cloning and USER Fusion: The Ideal Cloning Techniques for Small and Big Laboratories Hussam H. Nour-Eldin, Fernando Geu-Flores, and Barbara A. Halkier 14. Enrichment of Carotenoids in Flaxseed by Introducing a Bacterial Phytoene Synthase Gene Masaki Fujisawa and Norihiko Misawa 15. Metabolic Engineering by Plastid Transformation as a Strategy to Modulate Isoprenoid Yield in Plants Tomohisa Hasunuma, Akihiko Kondo, and Chikahiro Miyake 16. Engineering High Yields of Secondary Metabolites in Rubia Cell Cultures through Transformation with Rol Genes Victor P. Bulgakov, Yuri N. Shkryl, and Galina N. Veremeichik 17. Flow Cytometric Methods to Investigate Culture Heterogeneities for Plant Metabolic Engineering Vishal Gaurav, Martin E. Kolewe, and Susan C. Roberts 18. Phenylpropanoid Biosynthesis in Leaves and Glandular Trichomes of Basil (Ocimum basilicum L.) Cícero Deschamps and James E. Simon 19. Fusion with Fluorescent Proteins for Subcellular Localization of Enzymes Involved in Plant Alkaloid Biosynthesis Patrícia Duarte, Johan Memelink, and Mariana Sottomayor 20. Immunohistochemical Localization of a Putative Flavonoid Transporter in Grape Berries Elisa Petrussa, Enrico Braidot, Marco Zancani, Carlo Peresson, Alberto Bertolini, Sonia Patui, Valentino Casolo, Sabina Passamonti, Francesco Macri, and Angelo Vianello 21. Electrogenic Bromosulfalein Transport in Isolated Membrane Vesicles: Implementation ...